Composition for regulating immunological activity in intestine and use thereof

a technology of immunological activity and composition, applied in the direction of antibacterial agents, instruments, biocide, etc., can solve the problems of duox-mediated gut immunity activation, duox-mediated mucosal immunity defects, etc., and achieve the effect of preventing symptoms and efficiently using

Active Publication Date: 2016-06-02
SEOUL NAT UNIV R&DB FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0019]The composition of the present invention has effects for preventing symptoms caused by bacteria according to a mechanism which has been unknown in the art. Further, the present invention can be efficiently utilize

Problems solved by technology

However, the microbe-derived factors involved in the activation of DUOX-mediated gut immunity have not yet been determined.
However, the reason why gut immunity caused by DUOX is not activated by commensal microbes or the molecular mechanism causing immunolo

Method used

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  • Composition for regulating immunological activity in intestine and use thereof
  • Composition for regulating immunological activity in intestine and use thereof
  • Composition for regulating immunological activity in intestine and use thereof

Examples

Experimental program
Comparison scheme
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example 1

Fly Species and Rearing

[0176]In the present invention, the fly lineages including w1118 UAS-PLCβ-RFP (Ha et al., Nat Immunol 10, pp. 949-957, 2009), MEKKIUr3 (Inoue et al., Embo J 20, pp. 5421-5430, 2001), Drs-GFP, Dpt-LacZ (Jung et al., Biotechniques 30, pp. 594-598, 2001), imd1 (Lemaitre et al., Proc Natl Acad Sci USA 92, pp. 9465-9469, 1995), UAS-DUOX-RNAi (Ha et al., Science 310, pp. 847-850, 2005), norp7 (Ha et al., Nat Immunol 10, pp. 949-957, 2009), Gaq1 (Ha et al., Nat Immunol 10, pp. 949-957, 2009), esg-GAL4>UAS-GFP (Micchelli and Perrimon, Nature 439, pp. 475-479, 2006), upd3-GAL4>USA-GFP (Agaisse et al., Dev Cell 5, pp. 441-450, 2003), Su(H)Gbe-LacZ (Bray and Furriols, Curr Biol 11, pp. 217-221, 2001), and Da-GAL4 (Giebel et al., Mech Dev 63, pp. 75-87, 1997) were used. Transgenic flies containing 2XSTAT-GFP were used to monitor JAK-STAT pathway activation (Bach et al., Gene Expr Patterns 7, pp. 323-331, 2007).

[0177]Drosophila were reared at a temperature of 25° C. The co...

example 2

Bacterial Strains and Culture Conditions

[0178]Carotovora-15, a subspecies of Erwinia carotovora, was obtained from Bruno Lemaitre (Buchon et al., Cell Host Microbe 5, pp. 200-211, 2009b). Commensalibacter intestini A911T, Gluconobacter morbifer G707T, Acetobacter pomorum, Lactobacillus plantarum, and Lactobacillus brevis were isolated from the present inventors' laboratory fly stocks (Roh et al., Appl Environ Microbiol 74, pp. 6171-6177, 2008; Ryu et al., Science 319, pp. 777-782, 2008). Bacterial strains identified in humans were obtained from the gene bank of the Korea Research Institute of Bioscience and Biotechnology. The present inventors constructed two URA− mutant bacteria, E. carotovora-pyrE::Tn5 and G. morbifer-carA::Tn5, by Tn5-mediated random mutant libraries for E. carotovora and G. morbifer. Detailed methods for Tn5-mediated random mutagenesis and screening strategies are disclosed in Examples below and shown in FIGS. 6a to 6j and 12a to 12e. An E. carotovora-pyrE::Tn5-...

example 3

Production of Concentrated Bacterial Supernatants

[0179]E. carotovora were cultured in M9 minimal media. Because all of the commensal bacteria do not grow on M9 minimal media, various vitamins and essential amino acids were experimented with as disclosed in the previous reference (Foda and Vaughn, 1953). The present inventors confirmed that only C. intestini can grow on M9 minimal media with an additional 0.2 mg / mL of p-aminobenzoic acid, 1 mg / mL of pantothenic acid, and 0.2 mg / mL of nicotinic acid. In the late exponential phase, each bacterial supernatant (derived from approx. 105 bacterial cells) was obtained from centrifugation and filtering (pore size of 0.2 mM). Supernatants were lyophilized in Speedvac, and pellets were dissolved in 20 mL of distilled water. The sample was desalinized by adding 100% methanol in a volume 50 times more than that of the sample. Following the centrifugation, the sample was lyophilized in Speedvac, and dissolved in 20 mL of distilled water.

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Abstract

The present invention relates to an antimicrobial composition for uracil non-secretory (URA−) bacteria, comprising uracil, uracil precursors, uracil analogs, or uracil-secreting opportunistic pathogens as an active ingredient; and a composition for preventing cell damage caused by uracil-secretory (URA+) bacteria, comprising ROS scavengers as an active ingredient. The present invention also relates to food or feed for antimicrobial activity and preventing cell damage, comprising said composition; a method for controlling dual oxidase (DUOX) activation, the generation of reactive oxygen species (ROS), or the level thereof; a method for producing cells, wherein DUOX activation, the generation of ROS, or the level thereof are controlled; and a method for providing information on gut pathogenicity in isolated bacteria. The composition of the present invention has effects for preventing symptoms caused by bacteria according to a mechanism which has been unknown in the art. Further, the present invention can be efficiently utilized for establishing prevention and treatment strategies for symptoms caused by bacteria later, as therapeutic strategies can be readily established by bacterial classification based on criteria that can be easily confirmed.

Description

TECHNICAL FIELD[0001]The present invention relates to an antimicrobial composition for uracil non-secretory (URA−) bacteria comprising uracil, uracil precursors, uracil analogs, or uracil-secreting opportunistic pathogens as an active ingredient; and a composition for preventing cell damage caused by uracil-secretory (URA+) bacteria, comprising ROS scavengers as an active ingredient. The present invention also relates to food or feed for antimicrobial activity and preventing cell damage, comprising said composition; a method for controlling dual oxidase (DUOX) activation, the generation of reactive oxygen species (ROS), or the level thereof; a method for producing cells, wherein DUOX activation, the generation of ROS, or the level thereof are controlled; and a method for providing information on gut pathogenicity in isolated bacteria.BACKGROUND ART[0002]The gut epithelia of most metazoan organisms encompass complex microbial communities that range from autochthonous bacteria to allo...

Claims

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Application Information

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IPC IPC(8): A61K31/513G06F17/30
CPCA61K31/513G06F17/30598A23L1/30A61K31/01A61K31/352A61K31/353A61K31/355A61K31/375A61K31/409A61K31/505A61K31/522A61K31/7072A61K45/06A23L33/10A61P31/04A61K31/506G06F16/285
Inventor LEE, WON JAE
Owner SEOUL NAT UNIV R&DB FOUND
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